CN112218546A

CN112218546A - Compositions comprising human milk oligosaccharides for improving, enhancing, promoting or modulating gabaergic function in the central nervous system

Info

Publication number: CN112218546A
Application number: CN201980037220.2A
Authority: CN
Inventors: J·豪泽; 闫健
Original assignee: Societe des Produits Nestle SA
Current assignee: Societe des Produits Nestle SA
Priority date: 2018-06-25
Filing date: 2019-05-21
Publication date: 2021-01-12
Anticipated expiration: 2039-05-21
Also published as: US20210268005A1; AU2019296553A1; BR112020023102A2; MX2020012348A; PH12020551823A1; CL2020002978A1; EP3809879A1; CN112218546B; WO2020001863A1

Abstract

The present invention relates to a nutritional composition comprising Human Milk Oligosaccharides (HMOs) having the effect of improving, enhancing, promoting or modulating gabaergic function in the CNS of a mammal, preferably a human infant or young child born between the age of 7 years. The composition may be an infant formula. The HMO may be 2FL, diFL, and/or LNT and/or LNnT or a combination thereof.

Description

Compositions comprising human milk oligosaccharides for improving, enhancing, promoting or modulating gabaergic function in the central nervous system

Background

The present invention relates generally to the field of neuronal health, neuronal protection and neuronal development. In particular, the present invention relates to a composition for supporting neurodevelopment, in particular for improving, enhancing, promoting or modulating the function of or affecting the neurotransmitter gamma-aminobutyric acid (GABA) in the Central Nervous System (CNS) and other related cognitive benefits, in particular for infants and young children, preferably for a target population of infants or young children born between the age of 7 years (preterm or term).

More particularly, the present invention relates to the administration OF Human Milk Oligosaccharides (HMOs) for improving, enhancing, promoting or modulating gabaergic function in the CNS, in particular the filtering OF information or the ability to focus or concentrate on mental or physical activity, optionally in combination with additional oligomeric prebiotics, in particular Fructooligosaccharides (FOS) Or Fructooligosaccharides (OF) and/or Bovine Milk Oligosaccharides (BMOS).

The CNS and in particular the brain drives cognitive functions. The cerebral cortex is the nervous tissue layer of the outermost brain layer of the mammalian brain, and plays a key role in information integration of attention, sensory cognition, higher-order cognition (executive function) and sensory input.

CNS development and maturation are highly complex biological phenomena involving a large number of physiological processes including, for example, the growth and differentiation of neurons and glial cells, the guidance and branching of neurons, and the establishment of inter-neuronal conduction (neural signals) through axonal growth and synaptogenesis.

Neuronal plasticity, defined as the ability of the brain to continually adjust its functional and structural organization to alter demand, is important for the maturation and adult function of the nervous system. It is essential for the brain to function properly and for cognition, learning and memory. Some neuronal markers required for or at least associated with these physiological processes, including proteins and neurotrophic factors, such as brain-derived neurotrophic factor (BDNF) [ Huang, e.j. and Reichardt, L.F. (2001); neurotropic proteins, leaves in neural Development and Function, Annu.Rev.Neurosci, 24: 677-736; musumeci, G.and Minichiello, L. (2011); BDNF-TrkB signalling in near learning from genetics to neural networks, Rev. Neurosci, 22(3): 303-15); [ Xiao, j.et al. (2009); the role of neuropilins in The regulation of myelin, Neurosignals,17: 265-; immunological markers for sexual events, neurogenesis, and neurogenesis with the aid of the adaptive events, Cell Tissue Res, 345(1) (1-19).

The CNS begins to develop early after pregnancy, throughout pregnancy, and continues to mature until early adulthood. In particular, structural maturation is primarily prenatal, while functional network maturation is primarily postpartum. In the case of a human fetus, the development of the cerebral cortex is late and persists for a long period of time.

In utero, there is a peak in neuronal/brain maturation and growth from week 30 of pregnancy in humans.

The development of gabaergic function (including the ability to filter information, particularly sensory information such as visual signals) is a key step in the development of cognitive function in mammals, particularly infants and young children. Although this development and improvement of gabaergic function is particularly important during the first months/year of life (where neuronal plasticity is highest), it also affects older individuals, adolescents and adults, or elderly or diseased individuals.

By definition, premature infants enter the world with still primitive brains, and indeed they exhibit very basic electrical brain activity in primary sensory areas of the cerebral cortex (those areas that sense touch, vision and hearing) and primary motor areas of the cerebral cortex. For these infants, post-partum progressive maturation of the brain is necessary to compensate for its lower brain maturation state at birth, this compensatory maturation being particularly important for the more complex brain part that mediates most of its emotional, social and cognitive maturation in the first years of life [ Lubsen, j.et al (2011); microscopic and functional connectivity in the collapsing preterm bridge, sensines in robotics, 35,34-43 ].

Premature infants are born at a crucial moment in brain structural and functional development and maturation, and therefore they miss brain development processes in utero. Premature infants are at risk of medical conditions including hemorrhagic and ischemic-hypoxic brain injury after birth, and of developmental problems (including cognitive deficits) later in life. It appears that the more young the baby is at birth and the lighter the birth weight, the higher this risk. Cognitive deficits associated with low Intelligence Quotient (IQ), inattention and low working memory, and problems with executive function may persist through school age and adolescence [ Talge, n.et al. (2010) [ Late-term birthday and its Association with Cognitive and society outward communications at 6 ages of age, petro 1131; van baby, A., et al. (2009). Functioning at school at 32 to 36weeks' standing at Petrics, 124, 251-; farooqi, A et al, (2011). Impact at 11 years of major adjacent semiconductors in children born extreme prediction. Pediatrics,127, e 1247-1257; nosarti, C.et al, (2010), neuro-atmospheric outclocks of prediction birth. Cambridge: Cambridge University Press ].

More generally, immature CNS development or delayed CNS maturation can be observed in infants such as:

premature infants, low birth weight infants (<2500g), very low birth weight infants (<1500g), ultra low birth weight infants (<1000g), and small for gestational age infants [ Allen, M.C. (2008); neuro-atmospheric amounts of preterm inputs, curr. opin neurol, 21(2):123-8 ].

Premature or term infants who experience intrauterine growth retardation (IUGR) during pregnancy due to any adverse events (maternal smoking, maternal medication, poor placental quality, abnormal placental location, maternal and fetal malnutrition, maternal stress/anxiety, etc.); [ Gregory, A.et al. (2008); intrauterine Growth reaction effects the Preterm Infant's Hippocampus, Pediatric Research,63(4): 438-443 ].

Any neonate and small infant that exhibits a neurological developmental delay following, for example, hypoxia-ischemia at Birth, post-partum complications, post-partum steroid therapy or any other adverse event (see, for example, Barrett, R.D. et al (2007); Dedescription and recovery: hypoxia and the stabilizing cream, Birth Defects Res.C. embryo Today,81: 163-76).

These infants are reported to have cognitive dysfunction and also dysfunction in their growth and development, indicating that they do not achieve optimal "catch-up" for the neurodevelopmental process. Immature or delayed maturation of the cerebral cortex can lead to delayed and/or impaired learning ability, information integration, sensory input processing, loss or dysplasia of high-level reasoning ability, executive function, concentration, attention, motor skills, and language. This can lead to behavioral problems, abnormally low mental capacity, and thus abnormally low mental performance.

It is generally observed that breast-fed preterm infants may result in improved neurological development compared to formula feeding. (see, e.g., Roz re et al, the adaptation testing party in top prediction mechanisms: correlation between edition testing, early weight gain and neurology evaluation based on results from two countries, EPIPAGE and LIFT. BMJ Open 2012; 2: e 000834).

In healthy people, it has been observed that breast-fed infants improve cognitive function and education in adulthood compared to formula feeding. (see, e.g., visual et al association between scientific research and interpretation, and innovative at 30 layers of a progressive biological from Brazil. Lancet 2015)

According to the present inventors, this tends to indicate that some of the nutrients present in human breast milk may be missing from traditional/universal synthetic formulas or delivered in sub-optimal amounts. There is a need to identify key differences between conventional formulas and human breast milk and adjust the synthetic formulas accordingly.

Behavioral and neurodevelopmental disorders associated with delayed maturation of the cerebral cortex, in particular pathological gabaergic functions, including attention deficit/hyperactivity disorder, spectrum disorders of autism, and schizophrenia.

Cognitive function in humans can be measured in clinical tests, and is age dependent; many such tests are known to pediatricians and children's developmental experts. There are development screening and neurodevelopmental tests for babies and infants such as, for example, the BSID-belite infant development scale, the blaketan newborn behavior assessment scale, the NEPSY-developmental neuropsychological test, and the griffis mental development scale. Cognitive ability tests for preschool and/or school-age children include PPVT (peabody photo vocabulary test), TONI-2 (non-verbal intelligence test-2), WPPSI (wechsler preschool child intelligence scale), and CPM (rewiny color graphical inferential test).

It is known that nutrients play an important Role in The maturation of neurons in The Brain (reviewed in Huppi, P.S. (2008); Nutrition for The Brain, Peadiatric Research,63(3): 229-.

The consequences of malnutrition may be irreversible and may include poor cognitive development, poor memory, poor educational properties, and thus poor future economic productivity. (see, e.g., Horton, R; (2008) The Lancet, Vol.371, Issue 9608, page 179; Laus, M.F. et al. (2011); Early post native protein-cloning map and verification: a review of human and animal students, int.J. environ. Res. public health.,8(2): 590-). 612).

While it is known that mother's breast milk provides optimal nutritional support for the developing brain, when breast feeding is not possible, it is desirable to provide synthetic nutritional compositions (such as infant formulas or follow-up infant formulas) that induce improvement or promote the development of optimal cognitive functions.

Oral intervention is therefore a suitable way to positively influence the development of the nervous system in order to ensure optimal development of cognitive functions, memory and mental performance in preterm or term neonates, infants, toddlers, children or adolescents or young animals.

However, little is known and has been demonstrated to date regarding the ability of a nutritional diet or nutritional composition to affect development or promote gabaergic function, and particularly in infants and young children.

Thus, there is a general need to promote and support the healthy establishment of cognitive functions, and there is a need to improve, enhance, promote or modulate gabaergic function in the CNS, in particular the filtering of information or the ability to focus or concentrate on mental or physical activity.

There is a need to avoid, prevent or compensate for attention deficit/hyperactivity disorder, autism spectrum disorder, and schizophrenia, especially in individuals in need thereof, and especially by promoting or modulating gabaergic function in the CNS.

There is a need to promote the development or improvement of such functions in preterm or non-preterm young individuals (in particular infants and young children, such as human infants or young children) between the ages of birth and 7 years.

There is a need to provide such nutritional interventions and/or prophylactic nutritional interventions in a form that is generally accepted by individual populations, particularly those that are the most vulnerable or needed in these populations. There is also a need to not cause adverse, side effects or negative effects in such populations.

There is a need to provide such solutions to a population of individuals in the simplest and most cost-effective manner, preferably not by using actual ingredients considered as drugs or medicaments, and preferably as part of the diet.

The invention is applicable to all mammals, including animals and humans, and is particularly applicable to infants, toddlers or young pets with the highest brain plasticity. Preferred target populations are human (preterm or term) infants or young children/pets born to between 7 years of age.

Disclosure of Invention

The inventors have surprisingly found that administration OF a specific oligosaccharide or a mixture OF specific oligosaccharides comprising HMOs alone or in combination with OF and/or BMOS is particularly effective in improving, enhancing, promoting or modulating gabaergic function in the CNS, or the filtering OF information or the ability to focus or concentrate on mental or physical activity. The administration of the oligosaccharide can be performed as part of a nutritional intervention or as a therapeutic intervention in an individual in need thereof.

Drawings

FIG. 1: the effect OF various diets OF piglets (HMO, BMO, HMO + BMO, OF + HMO) on hippocampal expression OF GABA receptor GABBR1 is shown. HMO, OF and BMO are as described in the examples.

FIG. 2: the effect OF various diets OF piglets (HMO, BMO, HMO + BMO, OF + HMO) on hippocampal expression OF GABA receptor GABBR2 is shown. HMO, OF and BMO are as described in the examples.

Detailed Description

Definition of

As used herein, the following terms have the following meanings.

The term "infant" refers to a child under the age of 12 months.

The term "young child" refers to a child between the ages of one and seven.

As used herein, the article "a" or "an" preceding an element or component is intended to mean a non-limiting number with respect to the instance (i.e., occurrence) of the element or component. Thus, "a" or "an" includes one or at least one, and the singular form of the word a or an element or component also includes the plural form unless the number clearly indicates the singular.

The term "human milk oligosaccharides" is abbreviated HMO and collectively refers to those oligosaccharides that are present in human milk and fall within the conventional definition adopted by anyone skilled in the art. HMOs include, but are not limited to, fucosylated oligosaccharides (such as 2 '-fucosyllactose, 3' -fucosyllactose, difucosyllactose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucohexaose, lacto-N-difucohexaose I, fucosyllacto-N-hexaose, fucosyllacto-N-neohexaose I, fucosyllacto-N-neohexaose II, difucosyllacto-N-hexaose I, difucosyllacto-N-neohexaose II, fucosyl-p-lacto-N-hexaose, fucosyllactose, di-N-hexaose I, di-fucosyllacto-N-neohexaose II, and mixtures thereof, And any combination thereof), N-5 acetylated oligosaccharides such as lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), and any combination thereof), and sialylated oligosaccharides.

The term "bovine milk oligosaccharides" (abbreviated BMO) refers to those oligosaccharides which are present in bovine milk and fall within the conventional definition adopted by anyone skilled in the art. The BMO mixture used in the context of the present invention may for example be derived from bovine milk whey. Briefly, as described in the art, a bovine whey ultrafiltration permeate containing oligosaccharides such as 3 '-sialyllactose and 6' -sialyllactose and GOS can be demineralized by a combination of electrodialysis and ion exchange. BMOs may include, but are not limited to, fucosylated oligosaccharides (such as 2 '-fucosyllactose, 3' -fucosyllactose, difucosyllactose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucohexaose, lacto-N-difucohexaose I, fucosyllacto-N-hexaose, fucosyllacto-N-neohexaose I, fucosyllacto-N-neohexaose II, difucosyllacto-N-hexaose I, difucosyllacto-N-neohexaose II, fucosyl-p-lacto-N-hexaose, fucosyllactose, di-N-hexaose I, di-fucosyllacto-N-hexaose II, and mixtures thereof, And any combination thereof), N-5 acetylated oligosaccharides such as lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), and any combination thereof), and sialylated oligosaccharides.

As used herein, the term "fructooligosaccharide" (abbreviated OF) refers to fructooligosaccharides (i.e. fructooligosaccharides) having a degree OF polymerization OF 2 to 10, e.g. a degree OF polymerization OF 2 to 8. OF may also be referred to as fructooligosaccharides (Fructo-Oligo-Saccharides, abbreviated FOS) or short chain fructooligosaccharides (abbreviated scFOS). In the present disclosure, the terms OF, FOS, scFOS have the same meaning and are used interchangeably.

Inulin containing long-chain polymers is specifically excluded from the definition OF the present invention OF. OF is distinguishable from inulin by its degree OF polymerization (inulin has much longer chains).

FOS/scFOS/OF are generally commercially available, for example under the trade name ORAFTI Oligofructose from Beneo GmbH (Mannheim, Ger)many) (e.g. composition of

P95) were obtained commercially.

The term "sialylated oligosaccharide" refers to an oligosaccharide having one or more sialic acid residues.

The term "fucosylated oligosaccharide" refers to an oligosaccharide having one or more fucose residues.

The term "sn-2 palmitate" as used herein means that palmitic acid is bonded to triglycerides at their sn-2 position.

Gamma aminobutyric acid or "GABA" is generally known as an inhibitory neurotransmitter in the CNS, and its function to inhibit neuronal activity involves, for example, synchronizing neuronal networks in the hippocampus. The term "gabaergic" means "involving or affecting the neurotransmitter GABA". For example, if a synapse uses GABA as its neurotransmitter, the synapse is GABA energy. In another example, gabaergic neurons can produce GABA. In another example, a substance is gabaergic if it is capable of eliciting an effect via interaction with the GABA system (such as by stimulating or blocking neurotransmission). In another example, a gabaergic or gabaergic agent is any chemical substance capable of altering the effect of GABA in the body or brain. Classes of gabaergic drugs include, but are not limited to, GABA receptor agonists, GABA receptor antagonists, and GABA reuptake inhibitors. A specific exemplary GABA receptor is the GABA type B receptor (GABBR).

As used herein, the term "gabaergic function in the Central Nervous System (CNS)" relates to any gabaergic function exerted by or imposed on any cell, part of a cell, cell receptor, system or circuit of cells comprised in the CNS, including but not limited to neurons and glial cells, such as astrocytes, oligodendrocytes, microglia and ependymal cells, e.g. related to gabaergic neurotransmission, GABA transport or GABA uptake. Gabaergic neurotransmission is essential for filtering afferent information in the CNS, thereby limiting, reducing or removing unwanted or unnecessary information. Thus, specific gabaergic functions in the CNS include functions that filter information or the ability to focus or concentrate on mental or physical activity. Several examples of information-filtered GABA-capable modulation have been demonstrated in the art. For example, hippocampal GABA content correlates with the ability to suppress the search for unwanted memory triggered by human reminders (Schmitz et al. nat Commun.2017,8(1): 1311). In the optic parietal cap of zebrafish, GABAergic interneurons are necessary for spatial filtration, allowing fish to target sufficiently small preys (Del Bene et al science 2010,330(6004): 669-673). In addition, striatal GABA levels occurring through astrocyte uptake have been shown to alter the message filtering properties of regulatable neurons (Goubard et al J Physiol 2011,589(9):2301 and 2319). Improving, enhancing, promoting or modulating gabaergic function in the CNS can be assessed, for example, by measuring the expression levels of known genes involved in the gabaergic system, such as gabaergic receptors or transporters. For example, hippocampal gene expression may indicate improvement, enhancement, promotion, or modulation of gabaergic function.

The term "nutritional composition" refers to a composition that provides nutrients to an individual. Such nutritional compositions are typically administered orally or intravenously, and they typically include a lipid or fat source and a protein source. Preferably, the nutritional composition is a complete nutritional blend that meets all or most of the nutritional needs of an individual (e.g., infant formula). The nutritional composition comprises a foodstuff.

The term "infant formula" refers to a foodstuff intended to be dedicated to the provision of infant nutrition during the first four to six months of life, and which may itself meet the diverse nutritional needs of such persons (subject to the provisions of article 1.2 of directive 91/321/EEC for infant and follow-up infant formulas awarded by the European Commission on 1991, 5/14).

The term "follow-on formula" refers to a foodstuff that is dedicated to supplying nutrition to infants over four months of age and constitutes the main liquid component of the diet that is gradually diversified for such persons.

The term "starter infant formula (starter infant formula)" refers to a foodstuff intended to be dedicated to the nutrition of an infant during the first four months of life.

Infant formulas, follow-on infant formulas and starter 1 infant formulas may be in liquid form, ready-to-use or concentrated, or in the form of a dry powder that can be reconstituted with the addition of water to form the formula. Such formulations are well known in the art.

The term "baby food" refers to a foodstuff intended to be dedicated to the nutrition of an infant during the first year of life.

The term "infant cereal composition" refers to a foodstuff intended to be dedicated to the nutrition of infants during the first year of life.

The term "growing-up milk" refers to milk-containing beverages suitable for the specific nutritional needs of young children.

The term "weaning period" refers to the period in which breast milk is replaced with other food in the infant's diet.

The term "synthetic mixture" refers to a mixture obtained by chemical and/or biological means, which mixture may be chemically identical to the mixture naturally occurring in mammalian milk.

The term "prebiotic" refers to a non-digestible carbohydrate that exerts a beneficial effect on the host by selectively stimulating the growth and/or activity of healthy bacteria, such as bifidobacteria (bifidobacteria) in the human colon (Gibson GR, Roberfroid mb. diagnostic modulation of the human collagen microbiota: interconnecting the consortium. j nurr.1995; 125: 1401-12).

The term "probiotic" refers to a microbial cell preparation or microbial cell component that has a beneficial effect on the health or wellness of the host. (Salminen S, Ouwenand A. Benno Y. et al, "Probiotics: how shouldold the be defined" Trends Food Sci. Technol.1999: 10107-10).

All percentages are by weight unless otherwise indicated.

When the amount of an ingredient is provided as the weight of the ingredient per weight of the powdered nutritional composition, it is also intended that the present invention also includes corresponding amounts in liters to account for a dilution factor of 130g/L for the dry powdered nutritional composition (or otherwise indicated in the dilution specification).

Human milk oligosaccharides：

HMOs are collectively the third largest solid component of human milk after lactose and fat. HMOs are typically composed of lactose at the reducing end and a carbohydrate core at the non-reducing end, which typically contains fucose or sialic acid. More than one hundred lacto-oligosaccharides have been isolated and characterized, however, these oligosaccharides represent a very small fraction of the total number of oligosaccharides that have not yet been characterized.

Infant formulas have been developed using HMO ingredients such as fucosylated oligosaccharides, in particular 2FL, lacto-N-tetraose, lacto-N-neotetraose or sialylated oligosaccharides for different purposes.

EP0975235B1 from Abbott Laboratories describes synthetic nutritional compositions comprising one or more HMOs, wherein the HMOs in the composition are selected from the group consisting of eight HMOs (3-fucosyllactose, lacto-N-fucopentaose III, lacto-N-fucopentaose II, difucosyllactose, 2' -fucosyllactose, lacto-N-fucopentaose I, lacto-N-neotetraose, and lacto-N-fucopentaose V), wherein the composition is intended for use in the context of normal healthy infants, children, adults, or individuals with specific needs, such as those attendant with certain pathological conditions. EP0975235B1 indicates that oligosaccharides protect infants in general against viral and bacterial infections of the respiratory, gastrointestinal and genitourinary tracts.

In one embodiment, the composition of the invention comprises HMOs selected from the group consisting of: fucosylated oligosaccharides such as 2' -fucosyllactose (2FL), sialylated oligosaccharides and/or N-acetyl-lactosamines such as lacto-N-neotetraose (LNnT) or lacto-N-tetraose (LNT) or combinations thereof. Compositions comprising 2FL are exemplified herein experimentally.

N-acetyl-lactosamine

In some embodiments, the composition of the invention comprises at least one N-acetyl-lactosamine. This means that the composition according to the invention comprises N-acetyl-lactosamine and/or oligosaccharides comprising N-acetyl-lactosamine. Suitable oligosaccharides comprising N-acetyl-lactosamine include lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

Thus, in a preferred embodiment, N-acetyl-lactosamine is preferably selected from the group comprising: lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

LNT and LNnT can be chemically synthesized using an enzymatic transfer method (e.g., as described in U.S. Pat. No. 5,288,637 and WO 96/10086) using a glycosyltransferase to transfer the sugar unit of the donor moiety to the acceptor moiety. Alternatively, LNTs and lnnts can be prepared by chemically converting a keto-hexasaccharide (e.g., fructose) that is free or bound to an oligosaccharide (e.g., lactulose) to N-acetylhexamine or an oligosaccharide comprising N-acetylhexamine, such as Wrodnigg, t.m.; stutz, A.E, (1999) Angew. chem. int. Ed.38: 827-828. The N-acetyl-lactosamine prepared in this way may then be transferred to lactose as acceptor moiety.

Preferably, the composition according to the invention comprises 0.1g to 3g N-acetyl-lactosamine per 100g of the composition on a dry weight basis. Preferably, it comprises 0.1g to 3g of LNnT per 100g of the composition on a dry weight basis.

In one embodiment the nutritional composition according to the invention comprises N-acetyl-lactosamine, preferably selected from the group comprising: lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

Sialylated oligosaccharides

In some embodiments, the composition according to the invention may comprise one or more sialylated oligosaccharides.

Sialylated oligosaccharides include all sialylated HMOs, including 3 '-sialyllactose and 6' -sialyllactose. Preferably, both 3 '-sialyllactose and 6' -sialyllactose are present in the composition. In this embodiment, the ratio between 3 '-sialyllactose and 6' -sialyllactose is preferably in the range of from 100:1 to 1:100, more preferably from 10:1 to 1:10, even more preferably from 5:1 to 1: 2.

The 3 '-and 6' -forms of sialyllactose can be isolated from natural sources, such as animal milk, using chromatographic techniques or filtration techniques. Alternatively, sialylated oligosaccharides may also be prepared by biotechnological means, by enzyme-based fermentation techniques (recombinant or natural enzymes), by chemical synthesis or by microbial fermentation techniques, using specific sialyltransferases or sialidases, neuraminidases. In the latter case, the microorganism may express its native enzyme and substrate, or may be engineered to produce the corresponding substrate and enzyme. A single microbial culture or a mixed culture may be used. The formation of sialyloligosaccharides can start with an acceptor substrate having initially an arbitrary Degree of Polymerisation (DP), starting with DP ═ 1. Alternatively, sialyllactose may be produced by chemical synthesis from lactose and free N' -acetylneuraminic acid (sialic acid). Sialyllactose is also commercially available from, for example, Kyowa Hakko Kogyo, japan.

Preferably, the composition according to the invention comprises from 0.05g to 10g, more preferably from 0.1g to 5g, even more preferably from 0.1g to 2g sialylated oligosaccharide per 100g of the composition by dry weight.

In one embodiment, the nutritional composition according to the invention comprises sialylated oligosaccharides, which are preferably selected from the group comprising: 3 '-sialyllactose and 6' -sialyllactose. More preferably, the composition comprises both 3 '-sialyllactose and 6' -sialyllactose, the ratio between 3 '-sialyllactose and 6' -sialyllactose preferably being in the range of from 100:1 to 1:100, more preferably from 10:1 to 1:10, even more preferably from 5:1 to 1: 2.

Fucosylated oligosaccharide

The composition according to the invention may comprise one or more fucosylated oligosaccharides. Preferably, the fucosylated oligosaccharide consists of or comprises 2' -fucosyllactose (2-FL).

The fucosylated oligosaccharide may be selected from the group comprising: 2' -fucosyllactose, 3-fucosyllactose, Difucosyllactose (DiFL), lacto-N-fucopentaose (this means lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III and lacto-N-fucopentaose V), lacto-N-difucohexaose I, fucosyllacto-N-hexaose, difucosyllacto-N-hexaose I and difucosyllacto-N-neohexaose II. Particularly preferred fucosylated oligosaccharides are 2' -fucosyllactose (2-FL) or DiFL.

The fucosylated oligosaccharides can be isolated from natural sources (such as animal milk) using chromatographic techniques or filtration techniques. Alternatively, fucosylated oligosaccharides can also be prepared by biotechnological means by using enzyme (recombinant or natural) based fermentation techniques or microbial fermentation techniques using specific fucosyltransferases and/or fucosidases. In the latter case, the microorganism may express its native enzyme and substrate, or may be engineered to produce the corresponding substrate and enzyme. Single microbial cultures and/or mixed cultures may be used. Fucosylated oligosaccharides can be formed starting from acceptor substrates initially having any Degree of Polymerization (DP), starting from DP ═ 1. Alternatively, fucosylated oligosaccharides can be prepared by chemical synthesis from lactose and free fucose. Fucosylated oligosaccharides can also be obtained from, for example, Kyowa Hakko Kogyo, Japan.

Preferably, the composition according to the invention comprises between 0.1g and 3g fucosylated oligosaccharide, most preferably 2FL, by dry weight per 100g of the composition.

In one embodiment, the nutritional composition according to the invention comprises fucosylated oligosaccharides, preferably selected from the group comprising: 2 '-fucosyllactose, 3-fucosyllactose, difucosyllactose, lacto-N-fucopentaose (this means lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III and lacto-N-fucopentaose V), lacto-N-difucohexaose I, fucosyllacto-N-hexaose, difucosyllacto-N-hexaose I and difucosyllacto-N-neohexaose II, and preferably the fucosylated oligosaccharide is 2' -fucosyllactose (2-FL).

Additional prebiotics

In addition to the essential oligosaccharides claimed in this patent, the composition of the invention may also comprise at least one or one additional prebiotic, typically in an amount of from 0.3% to 10% by weight of the composition.

Prebiotics are generally non-digestible in the sense that they are not broken down and absorbed in the stomach or small intestine, and thus remain intact when they pass through the stomach and small intestine to the colon, where they are selectively fermented by beneficial bacteria.

In some embodiments, the composition according to the invention may comprise fructooligosaccharides (OF). An example OF such OF is the commercial ingredient OF Beneo GmbH (Mannheim, Germany)

In some embodiments, the prebiotic of the compositions of the present invention comprises other fructo-oligosaccharides (FOS) or/and galacto-oligosaccharides (GOS). Combinations of prebiotics may be used, such as 90% GOS combined with 10% short chain fructooligosaccharides (such as those sold under the trademark BENEO-Orafti corporation) "

oligofructise "(see http:// www.beneo-oratti. com/Our-Products/oligofructise) (formerly: oligofructise;" product of the invention)

) Or 90% GOS in combination with 10% inulin (such as is known under the trademark "by BENEO-Orafti corporation)"

Inulin "(see http:// www.beneo-orafti. com/Our-Products/Inulin) (formerly known as Inulin)

)). A further prebiotic combination was 70% short chain fructo-oligosaccharides combined with 30% inulin, which resulted from

Products sold under the trademark "Prebio 1And (5) preparing the product.

In one embodiment, the nutritional composition according to the invention comprises prebiotics selected from the following list: bovine milk oligosaccharides, inulin, xylo-oligosaccharides, polydextrose, or any combination thereof.

In one embodiment, the nutritional composition according to the invention comprises Bovine Milk Oligosaccharides (BMO). The BMO may comprise an oligosaccharide that is an N-acetylated oligosaccharide, a galactooligosaccharide, a sialylated oligosaccharide, a fucosylated oligosaccharide, or a combination thereof.

Probiotics

The composition of the invention may also comprise at least one probiotic. Non-limiting examples of probiotics include: bifidobacterium (Bifidobacterium), Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Enterococcus (Enterococcus), Streptococcus (Streptococcus), Kluyveromyces (Kluyveromyces), saccharomyces (saccharomyces), Candida (Candida), in particular selected from the group consisting of: bifidobacterium longum (Bifidobacterium longum), Bifidobacterium lactis (Bifidobacterium lactis), Bifidobacterium animalis (Bifidobacterium animalis), Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium infantis (Bifidobacterium infantis), Bifidobacterium adolescentis (Bifidobacterium adolescentis), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), Lactobacillus paracasei (Lactobacillus paracasei), Lactobacillus salivarius (Lactobacillus salivarius), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus plantarum (Lactobacillus lactis), Lactobacillus rhamnosus (Lactobacillus rhamnoides), Lactobacillus johnsonii (Lactobacillus johnsonii), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus plantarum (Lactobacillus), Lactobacillus lactis (Lactobacillus), Lactobacillus plantarum (Saccharomyces cerevisiae), Lactobacillus plantarum (Lactobacillus), Lactobacillus plantarum), Lactobacillus species (Lactobacillus), Bacillus lactis (Lactobacillus), Bacillus lactis (Bacillus lactis), Bacillus lactis (Bacillus lactis), Bacillus lactis, or mixtures of the following preferably selected from the group: bifidobacterium longum NCC3001(ATCC BAA-999), Bifidobacterium longum NCC2705(CNCM I-2618), Bifidobacterium longum NCC490(CNCM I-2170), Bifidobacterium lactis NCC2818(CNCM I-3446), Bifidobacterium breve strain A, Lactobacillus paracasei NCC2461(CNCM I-2116), Lactobacillus johnsonii NCC533(CNCM I-1225), Lactobacillus rhamnosus GG (ATCC53103), Lactobacillus rhamnosus NCC4007(CGMCC 1.3724), enterococcus faecium SF 68(NCC2768, NCIMB10415), and combinations thereof.

In one embodiment, the probiotic is a probiotic bacterial strain, preferably a bifidobacterium and/or a lactobacillus. Suitable probiotic bacterial strains include Lactobacillus rhamnosus (Lactobacillus rhamnosus) ATCC53103, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus paracasei (Lactobacillus paracasei) CNCM I-2116, Lactobacillus johnsonii (Lactobacillus johnsonii) CNCM I-1225, Lactobacillus delbrueckii Technologies Limited (BLIS Technologies Limited, New Zealand) Streptococcus salivarius DSM 13084, Bifidobacterium lactis anshan corporation (Lactobacillus sansentense, Denmark) Bifidobacterium lactis (Bifidobacterium) Bifidobacterium dolicCM 1-3446, Bifidobacterium longum Bifidobacterium A (Bifidobacterium longum Biocide) ATCC 3446, Bifidobacterium longum Bigenum A-33, Bifidobacterium longum Bigenum A-33, Bifidobacterium longum Bigenum Bignoni-b 12 Bifidobacterium breve sold under the trademark M-16V by Morinaga, Bifidobacterium infantis sold under the trademark Bifantis by Procter & GambIe Co, and Bifidobacterium breve sold under the trademark R0070 by the institute of Rosell biol, Canada.

Preferably, the composition according to the invention comprises 10e3 to 10e12cfu of probiotic bacterial strains, more preferably 10e7 to 10e12cfu of probiotic bacterial strains per 1g of the composition on a dry weight basis.

In one embodiment the nutritional composition comprises a nutritional composition selected from the group consisting of Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus salivarius (Lactobacillus salivarius), Lactobacillus rhamnosus, Lactobacillus paracasei (Lactobacillus paracasei), Lactobacillus casei (Lactobacillus casei), Lactobacillus johnsonii, Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus lactis (Lactobacillus lactis), Lactobacillus delbrueckii (Lactobacillus delbrueckii), Lactobacillus helveticus (Lactobacillus helveticus), Lactobacillus bulgaricus (Lactobacillus bulgaricus), lactococcus lactis (Lactococcus lactis), Lactococcus lactis diacetyl subspecies lactis (Lactococcus diacetylactis), Lactococcus lactis cremoris (Lactococcus cremoris), Streptococcus salivarius, Streptococcus thermophilus (Streptococcus thermophilus), Bifidobacterium lactis, Bifidobacterium animalis (Bifidobacterium animalis), Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, or Bifidobacterium adolescentis, or a probiotic bacterial strain of any mixture thereof.

Target population

In one embodiment, the invention is directed to a mammalian subject. Preferably, the mammal is a human, or a companion animal such as a dog or cat.

In a more preferred embodiment, the invention is directed to a young mammal, such as a young human, for example an infant (e.g. 0 to 6 months or 0 to 12 months), a toddler (e.g. 1 to 3 years or 1 to 7 years), or a puppy (e.g. puppy) or kitten. In an even more preferred embodiment, the mammal is an infant or a young child. Without being bound by theory, young mammals have high brain plasticity and brain (or brain connectivity) development and benefit most of the benefits of the present invention. It is also contemplated that the present invention may be directed to a subpopulation (fragile young mammals; aged or semi-aged mammals) having a particular need or at a stage of recovery for a particular window of nutritional intervention (e.g., children such as 6 months to 3 years, 3 months to 18 months). In one embodiment, the invention is particularly directed to individuals in need of avoiding, preventing or compensating for attention deficit/hyperactivity disorder, autism spectrum disorder, and schizophrenia (e.g., premature or mammals with suboptimal growth or development). In one embodiment, the present invention is particularly directed to individuals suffering from a health state or disease associated with gabaergic function, in particular a psychiatric or neurological disorder associated with gabaergic function, such as the filtering of information or the ability to focus or concentrate on mental or physical activity. Such diseases may include psychiatric or neurological disorders such as attention deficit/hyperactivity disorder, autism spectrum disorder, and schizophrenia.

According to a preferred embodiment, the composition according to the invention is for use in healthy infants and/or healthy young children. In one embodiment, the invention is of particular relevance in fragile infants, preterm infants and/or individuals with birth weights below normal at birth and/or infants with intrauterine growth retardation. Preferred periods of use are those periods of fastest development in memory and/or those periods of development in brain connectivity.

The present composition is directed to infants and/or young children 7 years or less, preferably 3 years or less, most preferably less than 1 year of age. In one embodiment, the composition is intended for infants of 6 months or less. In embodiments of the invention, the composition is used during the first 6 months of life, the first 1 year of life, the first 3 years of life, the first 7 years of life and/or during a period of recovery after disease or low development.

Nutritional composition

The nutritional composition according to the invention is preferably a synthetic nutritional composition. The composition of the invention may for example be a starter 1 infant formula, an infant formula, a baby food, an infant cereal composition, a follow-up formula or a growing-up milk, and preferably the composition is a starter 1 infant formula. The composition according to the invention may also be used before and/or during the weaning period. In one embodiment, the nutritional composition may be a complete nutritional composition or supplement for aged, elderly or fragile people.

The composition according to the invention may be a complete composition providing 100% or most of the nutritional needs of the target population (e.g. in terms of caloric needs; or in terms of vitamin or mineral needs, in terms of protein, lipid or carbohydrate needs). Alternatively, the composition of the present invention may be used as a supplement to be consumed in addition to a regular diet). However, in this case, the dosage and total consumption of the composition are adapted to provide the claimed benefits in terms of filtering of the information (e.g., proportional to caloric load and individual caloric demand).

The use of the composition of the invention may encompass the case where the composition is a supplement, preferably provided in unit dose form. In one embodiment, the composition is a supplement for human breast feeding.

The composition may be in the form of a powder composition, e.g. intended to be diluted with water or mixed with milk (e.g. human breast milk), or ingested in powder form. In one embodiment, the composition of the invention is in liquid form; ready-to-drink or diluted in water or mixed with milk (e.g., human breast milk).

The composition according to the invention may also comprise a protein source, preferably in an amount of less than 2.5g/100kcal, or less than 2.0g/100kcal, even more preferably in an amount of less than 1.8g/100 kcal. In one embodiment, the protein content is less than 1.6g/100 kcal. The type of protein is considered to be immaterial to the present invention, provided that the minimum requirements for essential amino acid content are met and satisfactory growth is ensured. Thus, protein sources based on whey, casein and mixtures thereof may be used, as may protein sources based on soy. For whey proteins of interest, the protein source may be based on acid whey or sweet whey or mixtures thereof, and may contain alpha-lactalbumin and beta-lactoglobulin in any desired proportions.

The composition according to the invention typically comprises a carbohydrate source. This is particularly preferred in case the nutritional composition of the invention is an infant formula. In this case, any carbohydrate source commonly found in infant formulas may be used, such as lactose, sucrose, maltodextrin, starch and mixtures thereof, but the preferred carbohydrate source is lactose.

The composition according to the invention typically comprises a lipid source. This is particularly relevant in case the nutritional composition of the invention is an infant formula. In this case, the lipid source may be any lipid or fat suitable for use in infant formulas. Preferred fat sources include palmitoleic acid, high oleic sunflower oil, and high oleic safflower oil. The essential fatty acids linoleic and alpha-linolenic acid may be added, as well as small amounts of oils containing high quality preformed arachidonic acid and docosahexaenoic acid, such as fish oils or microbial oils. The ratio of n-6 fatty acids to n-3 fatty acids of the fat source is preferably from about 5:1 to about 15: 1; for example from about 8:1 to about 10: 1.

The composition of the present invention also preferably contains all vitamins and minerals that are considered essential to the daily diet in nutritionally significant amounts. The minimum requirements for certain vitamins and minerals have been determined. Examples of minerals, vitamins and other nutrients optionally present in the compositions of the present invention include vitamin a, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, folic acid, inositol, niacin, biotin, pantothenic acid, choline, calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chlorine, potassium, sodium, selenium, chromium, molybdenum, taurine and l-carnitine. The minerals are typically added in salt form. The presence and amounts of particular minerals and other vitamins will vary depending on the target population.

If necessary, the composition of the present invention may contain emulsifiers and stabilizers such as soybean, lecithin, citric acid monoglyceride and citric acid diglyceride, and the like.

The compositions of the present invention may also contain other substances that may have a beneficial effect, such as lactoferrin, nucleotides, nucleosides, and the like.

In one embodiment, the composition of the invention (especially in the form of an infant formula) comprises about 1.8g to about 2.2g total protein per 100kcal, for example about 1.8g to about 2.1g, or about 1.9g to about 2.1g protein per 100kcal, optionally wherein about 0.3g/100kcal to about 0.4g/100kcal of protein is alpha-lactalbumin. The infant and follow-on infant formulas of the present invention may be in the form of a ready-to-eat liquid, or may be a liquid concentrate or powdered formula that can be reconstituted into a ready-to-eat liquid by the addition of an amount of water that results in and conforms to the follow-on infant formula containing all the ingredients required by the U.S. or european union laws (including but not limited to certain vitamins, minerals and essential amino acids). It may also contain nucleotides (such as CMP, UMP, AMP, GMP and IMP), lutein, zeaxanthin and other ingredients known in the art.

In one embodiment of the invention, the nutritional composition is a pet food (e.g. for dogs or cats or puppies or kittens).

Effect and use of the compositions of the invention

The present invention relates to improving, enhancing, promoting or modulating gabaergic function in the CNS of an individual. This may include improving, enhancing, facilitating or adjusting the filtering of information or the ability to focus on mental or physical activity.

Without being bound by theory, in one aspect of the invention, it is believed that such improvement, enhancement and/or promotion is associated with the effect of the oligosaccharides of the invention on the gut microbiota. Oligosaccharides such as HMO (e.g., 2FL or LNnT), BMO or OF are only slightly digested in the small intestine and therefore most are available for fermentation by the microbiota in the colon. Indeed, HMOs (e.g. 2FL or LNnT), BMOs or OF have been shown to promote the growth OF bifidobacterium enterobacter, as reflected by an increase in such bacteria in the stool OF infants fed oligosaccharide supplement formulas (with HMOs (e.g. 2FL or LNnT), BMOs or OF). It appears that both oligosaccharides and probiotics are associated with beneficial alterations in GABA receptor expression in the brain. Without wishing to be bound by any theory, due to the high presence of GABA in some bacterial species (e.g., lactobacillus and bifidobacterium), such probiotic species can act as delivery mediators of GABA, and their ingestion can exert a local effect on the gastrointestinal system that ultimately regulates the host neurophysiology (Lyte, BioEssays 2011,33, 574-. Considering that both prokaryotes and eukaryotes synthesize GABA by decarboxylating glutamate via glutamate decarboxylase (GAD), this linkage appears even stronger, and analysis of items from the human microbiome indicates that the gene encoding GAD is present in the human microbiome (Mazzoli, r., and pessinone, E. (2016.). front. microbiol.7, 1934; Pokusaeva et al, neuroastroenol.motil.2017, 29(1) doi: 10.1111/nmo.12904).

In short, the nutritional compositions of the present invention can be used to improve, enhance, promote or modulate gabaergic function through its effects on the gut microbiota and gut brain axis, which has been shown to affect many aspects of brain function. It is also envisaged that improving, enhancing, promoting or modulating gabaergic function may be associated with an increase in the concentration of sialic acid (Neu5Ac) in the brain of the individual mediated by a differential metabolic pathway affected by the gut microbiota. Without being bound by theory, in one embodiment of the invention, HMOs act synergistically with probiotics and endogenous microbiota to optimally affect such metabolic pathways.

In another aspect, the underlying effect of the invention may be related to altered gene expression of a gabaergic gene, in particular a GABA receptor or GABA transporter, in the CNS of the individual. In one embodiment, alteration refers to an increase or decrease in the level of gene expression measured at the mRNA or protein level as compared to a control. In one embodiment, gene expression refers to hippocampal gene expression of an individual.

In another aspect, the underlying effects of the invention may be related (without being bound by theory) to enhancing neuroplasticity in the brain of an individual, and/or by enhancing neurodegeneration, neurogenesis, axonal sprouting, myelination, and/or maturation in the brain of the individual.

Gabaergic function, particularly the ability of information to filter or focus on mental or physical activity, represents an important aspect of brain function and development and is critical to the overall cognitive development of an individual. It affects especially the processing, classification and management of external signals, learning capabilities and spatial orientation.

In one embodiment of the invention, the gabaergic function is the ability to filter information such as incoming sensory information. Filtering information is crucial for social and cognitive development, especially for young individuals. In one embodiment of the invention, the gabaergic function is the ability to focus or concentrate on mental or physical activity.

Thus, in one aspect, the present invention relates to (or is defined by) the use of a nutritional composition comprising HMOs for improving, enhancing, promoting or modulating gabaergic function in the CNS, in particular the ability to filter information or the ability to focus or concentrate on mental or physical activity of a mammal.

In one aspect, the present invention relates to the use of (or is defined by) a nutritional composition comprising HMOs as described herein for avoiding, preventing or compensating for attention deficit/hyperactivity disorder, autism spectrum disorder and schizophrenia, particularly in an individual in need thereof, and particularly by promoting or modulating gabaergic function in the CNS.

In one aspect, the present invention relates to the use of (or is defined by) a nutritional composition comprising HMOs as described herein for improving, enhancing or modulating attention deficit/hyperactivity disorder, autism spectrum disorder and schizophrenia, and in particular by promoting or modulating gabaergic function in the CNS.

In one aspect, the present invention relates to (or is defined by) the use of a nutritional composition comprising HMOs as described herein for improving, enhancing or modulating gabaergic function in the CNS, in particular the ability to filter information or focus or concentrate on mental or physical activity.

In one embodiment, the present invention relates to the use of (or the invention is defined by) a nutritional composition as described herein, wherein the composition is an infant formula, a follow-on formula, a human milk fortifier, an adult milk, or a pet food.

In another aspect, the present invention relates to (or is defined by) an HMO for improving, enhancing, promoting or modulating gabaergic function, in particular the ability to filter information or focus or concentrate on mental or physical activity, in the CNS of an individual in need thereof. In a particular embodiment, the subject is a mammal, wherein the mammal is preferably a human, a cat or a dog, and wherein the human is preferably an infant or a young child.

In another aspect, the present invention relates to a nutritional composition (or defined by the nutritional composition) comprising HMOs for use in improving, enhancing, promoting or modulating gabaergic function in the CNS, in particular the ability to filter information or focus or concentrate on mental or physical activity, in an individual in need thereof. In a particular embodiment, the individual is a mammal, preferably the mammal is a young mammal, human, dog or cat, more preferably a young human, dog or cat, most preferably an infant or a young child. The nutritional compositions may further comprise one or more of the optional ingredients described herein.

In another aspect, the invention relates to the use of HMOs for the manufacture of (or as defined by) a nutritional composition for improving, enhancing, promoting or modulating gabaergic function, in particular the ability to filter information or the ability to focus or concentrate on mental or physical activities, in the CNS of an individual in need thereof. In a particular embodiment, the individual is a mammal, preferably the mammal is a young mammal, human, dog or cat, more preferably a young human, dog or cat, most preferably an infant or a young child.

Dosage form

HMO: in one embodiment, HMO is present in a total amount of 0.01 to 50g/L, 0.1 to 10g/L, 0.3 to 5g/L, or 0.5 to 1g/L, or 0.25g/L or 0.5g/L or 1g/L or 1.5g/L or 2 g/L.

BMO: in one embodiment, the BMO is present in a total amount of 0.01 to 50g/L, 0.1 to 10g/L, 0.3 to 5g/L, or 0.5 to 1g/L, or 0.25g/L or 0.5g/L or 1g/L or 1.5g/L or 2 g/L.

fructooligosaccharide/FOS/OF: when present, the nutritional composition OF the invention may comprise 0.1 to 20g OF Fructooligosaccharides (OF) per 100g OF the composition on a dry weight basis, e.g. 1 to 6g or 3 to 5g OF Fructooligosaccharides (OF) per 100g OF the composition on a dry weight basis.

In one embodiment of the invention, the nutritional composition comprises an amount of fructooligosaccharides in the following ranges or amounts:

0.1 to 20g/L or 0.5 to 10g/L or 1 to 8g/L or 2 to 6g/L or 1.5g/L or 3g/L or 5g/L of a nutritional composition when the composition is in the form of a ready-to-eat liquid, or

When the composition is in powder form and is intended to be reconstituted into a diluted liquid form, 0.1 to 20G/L or 0.5 to 10G/L or 1 to 8G/L or 2 to 6G/L or 1.5G/L or 3G/L or 5G/L, or

When the nutritional composition is in the form of a concentrated composition intended to be diluted (2, 5, 10, 20, 50 or 100 times, respectively) into water or human breast milk or intended to be used directly in concentrated form, the above values are multiplied by 2, 5, 10, 20, 50 or 100, or

When the nutritional composition is in the form of a dry powder, 0.4g to 15g/100g of the nutritional composition powder, or 0.8 to 10g/100g, or 1 to 6g/100g, or 2 to 5g/100g or 2.1 to 4g/100g or 1.2g/100g or 2.3g/100g or 3.8g/100g or 4g/100g or 6g/100g of the nutritional composition powder.

In one embodiment, when the nutritional composition is in the form OF a dry powder, the OF content may be 0.07g to 3g/100kcal OF the nutritional composition powder, or 0.1 to 2g/100kcal, or 0.4 to 1.5g/100kcal, or 0.45 to 1g/100kcal or 0.45 to 0.75g/100kcal or 0.3g/100kcal or 0.4g/100kcal or 0.5g/100kcal or 0.75g/100kcal or 1g/100kcal OF the nutritional composition powder.

2 FL: the nutritional composition of the invention may comprise 0.02g to 10g of 2FL per 100g of the composition on a dry weight basis, for example 0.2g to 0.5g or 0.3g to 5g of 2FL per 100g of the composition on a dry weight basis.

In one embodiment, the nutritional composition comprises an amount of 2FL in the following ranges or amounts:

0.05 to 20g/L or 0.1 to 5g/L or 0.2 to 3g/L or 0.1 to 2g/L or 0.25g/L to 1g/L or 0.25g/L or 1g/L of a nutritional composition when the composition is in the form of a ready-to-eat liquid, or

When the composition is in powder form and is intended to be reconstituted into a diluted liquid form, 0.05 to 20g/L or 0.1 to 5g/L or 0.2 to 3g/L or 0.1 to 2g/L or 0.25g/L to 1g/L or 0.25g/L or 1g/L, or

When the nutritional composition is in the form of a dry powder, 0.04 to 1.5g/100g of the nutritional composition powder, or 0.08 to 1.2g/100g, or 0.1 to 1g/100g, or 0.2 to 0.8g/100g or 0.2g/100g or 0.4g/100g or 0.8g/100g or 1g/100g of the nutritional composition powder.

In one embodiment, when the nutritional composition is in the form of a dry powder, the 2FL content may be 0.01g to 0.3g/100kcal of the nutritional composition powder, or 0.02 to 0.2g/100kcal, or 0.04 to 0.15g/100kcal, or 0.02g/100kcal, or 0.04g/100kcal, or 0.07g/100kcal, or 0.15g/100kcal, or 0.3g/100kcal of the nutritional composition powder.

LNnT: in one embodiment, the nutritional composition may comprise 0.01g to 1g of LNnT per 100g of the composition on a dry weight basis, for example 0.1g to 0.25g or 0.15g to 0.5g of LNnT per 100g of the composition on a dry weight basis.

In one embodiment of the invention, the nutritional composition comprises an amount of LNnT in the following ranges or amounts:

0.02 to 10g/L or 0.05 to 2.5g/L or 0.1 to 1.5g/L or 0.05 to 1g/L or 0.12g/L to 0.5g/L or 0.12g/L or 0.5g/L or 1g/L of a nutritional composition when the composition is in the form of a ready-to-eat liquid, or

When the composition is in powder form and is intended to be reconstituted into a diluted liquid form, 0.02 to 10g/L or 0.05 to 2.5g/L or 0.1 to 1.5g/L or 0.05 to 1g/L or 0.12g/L to 0.5g/L or 0.12g/L or 0.5g/L or 1g/L, or

0.02g to 0.75g/100g of the nutritional composition powder, or 0.04g to 0.6g/100g, or 0.0.5g to 0.5g/100g, or 0.1g to 0.4g/100g or 0.1g/100g or 0.2g/100g or 0.25g/100g or 0.5g/100g or 1g/100g or 3g/100g of the nutritional composition powder when the nutritional composition is in the form of a dry powder.

In one embodiment, when the nutritional composition is in the form of a dry powder, the LNnT content may be 0.01g to 0.3g/100kcal of the nutritional composition powder, or 0.02 to 0.2g/100kcal, or 0.04 to 0.15g/100kcal, or 0.02g/100kcal, or 0.04g/100kcal, or 0.07g/100kcal, or 0.15g/100kcal, or 0.3g/100kcal of the nutritional composition powder.

In particular, when in the form of an infant formula, the compositions of the invention may comprise at least about 0.4g of fructooligosaccharides per 100kcal of fructooligosaccharides. In some embodiments, it comprises per 100kcal from about 0.4g to about 0.9g, from about 0.4g to about 0.7g, from about 0.4g to about 0.5g, from about 0.7g to about 0.8g, or from about 0.7g to about 0.9g fructo-oligosaccharides. The fructooligosaccharides have a degree of polymerization of 2 to 10. In one embodiment, at least 90% of the fructooligosaccharides have a degree of polymerization of 2 to 8.

In one embodiment, the nutritional composition comprises 2FL and LNnT, preferably in an amount of 1 g/L2 FL and 0.5g/L LNnT, or 0.5 g/L2 FL and 0.25g/L LNnT, or 0.25g/L to 2 g/L2 FL and 0.1 to 1g/L LNnT.

Method for preparing a nutritional composition

The nutritional compositions may be prepared in any suitable manner known in the art. For example, commercial infant formulas such as larger infant formulas may be used as base compositions to which the desired amount OF oligosaccharides (e.g., HMOs such as 2FL, LNnT, etc., OF, BMO), preferably in dry form, is added. Alternatively, the oligosaccharides may be added as dry or liquid ingredients to a liquid premix which will serve as a basis for preparing the nutritional composition of the present invention. The liquid mixture may then be dried by any conventional method.

For example, the nutritional composition may be prepared by blending together a protein source, a carbohydrate source (other than the oligosaccharide combination of the invention) and a fat source in suitable proportions. If used, the emulsifier may be added at this point. Vitamins and minerals may be added at this point, but are usually added at a later time to avoid thermal degradation. Any lipophilic vitamins, emulsifiers, etc. may be first dissolved in the fat source prior to blending. Water (preferably water subjected to reverse osmosis) may then be mixed in to form a liquid mixture. The water temperature is suitably in the range of about 50 ℃ to about 80 ℃ to assist in dispersing the ingredients. Commercially available liquefiers may be used to form the liquid mixture. If the final product is in liquid form, 3 '-sialyllactose (3' -SL) and 6 '-sialyllactose (6' -SL) are added at this stage. If the final product is in powder form, 3 '-sialyllactose (3' -SL) and 6 '-sialyllactose (6' -SL) may be added at this stage as well, if desired. The liquid mixture is then homogenized, for example, in two stages.

The liquid mixture may then be heat treated to reduce bacterial loads, for example by rapidly heating the liquid mixture to a temperature in the range of about 80 ℃ to about 150 ℃ for a duration of between about 5 seconds and about 5 minutes. This may be done by steam injection, autoclave or heat exchanger (e.g. plate heat exchanger). The liquid mixture is then cooled, for example by chilling, to about 60 ℃ to about 85 ℃. The liquid mixture may then be homogenized again, for example in two stages, wherein the pressure of the first stage is between about 10MPa and about 30MPa and the pressure of the second stage is between about 2MPa and about 10 MPa. The homogenized mixture may then be further cooled in order to add any heat sensitive components such as vitamins and minerals. It is now convenient to adjust the pH and solids content of the homogenized mixture. The homogenized mixture is transferred to a suitable drying apparatus (such as a spray dryer or freeze dryer) and converted to a powder. The moisture content of the powder should be less than about 5% by weight.

If a liquid composition is preferred, the homogenized mixture may be sterilized and then filled into suitable containers under aseptic conditions, or it may be filled into containers and then distilled.

Specific lipids

The compositions of the invention may comprise selected lipids having specific effects.

These lipids may in particular comprise DHA, ARA, linoleic acid or sphingomyelin, preferably in an amount suitable for delivering the actual brain health benefit and within the general regulatory requirements of the product type (e.g. WHO recommendations for infant formula; CODEX or european instructions for infant formula).

In some embodiments, the compositions of the present invention comprise a relatively high content of sn2 palmitate or sphingomyelin. These are associated with optimal brain performance and development and may act synergistically with the basic compounds of the compositions of the present invention.

While feeding an infant with a formula comprising a high percentage OF sn-2 palmitate (in the absence OF) helps promote the growth OF bifidobacteria in the colon, it is believed that the combination OF high sn-2 palmitate with fructooligosaccharides provides significantly superior bifidobacteria growth in the colon OF formula-fed infants. A significant reduction in the amount of potential pathogenic bacteria can also be achieved. It has been found that feeding an infant formula comprising high sn-2 palmitate comprising from about 3 to about 5g/L, or from about 0.4 to about 0.7g/100kcal, of fructooligosaccharides to an infant is more beneficial than feeding the same formula to an infant without the fructooligosaccharides. Without being bound by theory, this synergistic effect between OF and sn2 palmitate may also promote short-term memory, although in particular (but most likely not exclusively) its effect on the microbiota and bifidobacteria populations OF an individual. By "high Sn2 palmitate" is understood a composition where the palmitate has a high percentage of fatty acids in the Sn2 position of the triglycerides, preferably more than 33% of the fatty acids. Such ingredients are available under the trade name

(Loders Croklaan, Wormerveer, Netherlands) or

(Advanced Lipids AB, Karlshamn, Sweden, joint vision of AAK B.V. (Zaandijk, Netherlands) and Enzytec Inc, Morristown, USA).

Recent clinical studies in infants have shown that nutritional formulas containing at least one omega 6 fatty acid and at least one omega 3 fatty acid in a ratio of about 6 to about 1 increase the accretion of DHA in red blood cells and plasma. An approximately 6:1 equilibrium ratio of omega 6 fatty acids to omega 3 fatty acids may also provide long-term health benefits, including optimal brain and neurological development. This balance will be achieved by formulating the present invention with a vegetable oil fat source having an omega 6 fatty acid content (such as, for example, soybean oil and sunflower oil) and an omega-3 fatty acid content (for example, rapeseed, canola, linseed, chia, perilla or walnut). A unique fat blend with 5 different oils will be used to achieve a modified fat blend.

The following examples are presented to illustrate certain embodiments and features of the invention, but should not be construed as limiting the scope of the invention.

Examples

Example 1: nutritional compositions comprising HMO (2FL and/or LNnT)

The nutritional compositions OF the present invention comprising HMO 2FL and/or LNnT and OF are given in table 1 below. The composition is given by way of illustration only. The composition of table 1 may be an infant formula. Alternatively, it may be suitable for larger infant formulas.

TABLE 1：

Example 2: infant formula comprising HMO (2FL and/or LNnT), optionally BMO and/or fructo-Oligosaccharides (OF) Article (A)。

The nutritional compositions OF the invention comprising HMOs (2FL and/or LNnT) and optionally OF and/or BMOs are given in table 2 below. The composition is given by way of illustration only. The compositions of table 2 are infant formulas. Alternatively, it may be suitable for larger infant formulas.

Another example specific oligosaccharides of the invention were added to commercial NAN and/or mechano-gen (lactose) infant formulas (available from nestle, Switzerland) in the following amounts.

TABLE 2

Experimental data

In a piglet study, we studied the effect OF various oligosaccharides (e.g., 2FL) containing HMO on hippocampal gene expression in the presence or absence OF and/or BMOS. Thirty-six male piglets (12 per treatment group) were treated with either a control (CON, Purina ProNurse Livestock Milk Replacer), HMO (CON +1.5g/L HMO (consisting OF 1.0 g/L2 ' FL +0.5g/L LNnT), OF [ CON +5g/L OF ], OF + HMO [ CON +5g/L OF +1.0g/L HMO (2' FL), BMO (6.5g/L BMO), or HMO + BMO (1.0 g/L2 ' FL +0.5g/L LNnT +6.5g/L BMO) from 48 hours post partum until 33 days OF life.A thirty-six male piglet was treated with formulation space reserved for the addition OF a dietary test article.all diets contained 8.0g/L OF formulation space from PND 2-6 and PND 7-33, respectively, at a reconstitution BW 285mL and 325mL diet, day 32 or 33 days OF euthanasia, and hippocampal tissues were collected for analysis of mRNA expression.

The BMO mixture used in the formula is derived from bovine milk whey. Briefly, bovine milk whey ultrafiltration permeate containing oligosaccharides such as 3 '-sialyllactose and 6' -sialyllactose and GOS was demineralized by a combination of electrodialysis and ion exchange as described in the art.

Hippocampus gene expression

Approximately 20mg of hippocampal Tissue was introduced into lysis matrix D-tubes (MP Biomedicals, Santa Ana, California, USA), placed on ice, and 650. mu.L of lysis buffer (Agencourt RNAderived Tissue Kit, Beckman Coulter, Indianapolis, Indiana, USA) was added. Is formed by arranging a pipe in

(MP Biomedicals, Santa Ana, California, USA) at speed 6 for 2X 1 min, followed by Agen JuRNAdvance Tissue Kit (Beckman C)outer, Indianapolis, Indiana, USA) 400. mu.L of lysate was extracted as recommended by the manufacturer. Using Quant-iT^TMRiboGreen^TMRNA assay kit (Invitrogen, Carlsbad, California, USA) RNA was quantified on Spectramax M2(Molecular Devices, Sunnyvale, California, USA). Quantitative assessment of RNA was accomplished using a fragment analyzer 96(Advanced Analytical Technologies, inc., Ankeny, Iowa, USA) with a standard sensitivity RNA analysis kit (15 nt). Use the NanoString nCounter^TMThe system (NanoString Technologies inc., Seattle, Washington, USA) quantifies the relative mRNA copy number on 93 genes using 100ng RNA as the starting amount according to the manufacturer's instructions.

Results

Compared to the CON group, the HMO, BMO + HMO and OF + HMO diets showed differential effects on gene expression. Hippocampal mRNA expression of several key genes involved in the gabaergic system is shown, in fig. 1 GABA type B receptor subunit 1(GABBR1) and in fig. 2 GABA type a receptor β 2 subunit (GABRB 2). Administration OF HMOs, preferably in combination with prebiotics such as OF and/or BMOs, has the surprising ability to modulate the gabaergic system at the expression level.

Figure 1 shows that mRNA expression OF the GABBR1 receptor gene was maintained or even significantly increased compared to the control diet (CON) when HMO was also present in the test diet alone (2' FL + LNnt) or in combination with OF or BMO.

Figure 2 shows that mRNA expression of the GABRB2 receptor was specifically increased when HMOs were administered in combination with BMOs compared to the control diet.

It is to be understood that one or more embodiments of the present disclosure are non-limiting embodiments, and that the present disclosure is intended to cover modifications and equivalents of these embodiments.

Claims

1. It is to be understood that the following claims are provided for purposes of illustration only and are not intended to limit the scope of the inventive concepts described or otherwise contemplated herein in any way.

Use of a nutritional composition comprising Human Milk Oligosaccharides (HMOs) for improving, enhancing, promoting or modulating a) gabaergic function in the Central Nervous System (CNS) or b) the ability of information to be filtered or focused on mental or physical activity in a mammal, preferably a human infant or young child born between the ages of 7.

2. Use of a nutritional composition comprising Human Milk Oligosaccharides (HMOs) in a mammal, preferably a human infant or young child born between 7 years of age, for avoiding, preventing or compensating attention deficit/hyperactivity disorder, autism spectrum disorder and schizophrenia, preferably by promoting or modulating gabaergic function in the CNS.

3. The use of the nutritional composition according to claim 1 or 2, wherein the gabaergic function in the CNS comprises filtering of information or the ability to focus or concentrate on mental or physical activities.

4. Use of a nutritional composition according to any of claims 1 to 3, wherein the HMO is selected from the group consisting of: n-acetyl-lactosamine, sialylated oligosaccharides, fucosylated oligosaccharides or a combination thereof.

5. Use of the nutritional composition according to claim 4, wherein the HMO is a fucosylated oligosaccharide selected from the group consisting of: fucosylated oligosaccharide, 2 '-fucosyllactose, 3' -fucosyllactose, difucosyllactose, lacto-N-fucopentaose I, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose V, lacto-N-fucohexaose, lacto-N-difucohexaose I, fucosyllacto-N-hexaose, fucosyllacto-N-neohexaose I, fucosyllacto-N-neohexaose II, difucosyllacto-N-hexaose I, difucosyllacto-N-neohexaose II, fucosyl-p-lacto-N-hexaose, di-fucosyllacto-N-hexaose II, fucosyl-p-lacto-N-hexaose, And any combination thereof, an N-5 acetylated oligosaccharide such as lacto-N-tetraose (LNT), lacto-N-neotetraose (LNnT), and any combination thereof, and preferably the fucosylated oligosaccharide is selected from the group consisting of 2' -fucosyllactose (2-FL), diFL, LNnT, LNT, and any combination thereof.

6. Use of the nutritional composition according to claim 4, wherein the HMO is N-acetyl-lactosamine selected from the group consisting of: lacto-N-tetraose (LNT) and lacto-N-neotetraose (LNnT).

7. Use of the nutritional composition according to claim 4, wherein the HMO is a sialylated oligosaccharide, preferably selected from the group consisting of: 3 '-sialyllactose and 6' -sialyllactose, and more preferably wherein the composition comprises both 3 '-sialyllactose and 6' -sialyllactose, the ratio between 3 '-sialyllactose and 6' -sialyllactose is preferably in the range of from 100:1 to 1:100, more preferably from 10:1 to 1:10, even more preferably from 5:1 to 1: 2.

8. Use of a nutritional composition according to any of the preceding claims, wherein the HMO is present in an amount of from 0.01 to 50g/L, from 0.1 to 10g/L, from 0.3 to 5g/L or from 0.5 to 1g/L, or from 0.25g/L or from 0.5g/L or 1g/L or 1.5g/L or 2 g/L.

9. Use of a nutritional composition according to any of the preceding claims, further comprising a prebiotic, preferably selected from the list consisting of: bovine Milk Oligosaccharides (BMO), inulin, xylo-oligosaccharides, polydextrose, Fructooligosaccharides (FOS), Or Fructooligosaccharides (OF), or any combination thereof.

10. Use of the nutritional composition according to claim 9, wherein the BMOs comprise N-acetylated oligosaccharides, galactooligosaccharides, sialylated oligosaccharides, fucosylated oligosaccharides or a combination thereof.

11. Use of a nutritional composition according to any of the preceding claims, further comprising a probiotic, wherein the probiotic is preferably selected from the list consisting of: bifidobacterium (Bifidobacterium), Lactobacillus (Lactobacillus), Lactococcus (Lactococcus), Enterococcus (Enterococcus), Streptococcus (Streptococcus), Kluyveromyces (Kluyveromyces), saccharomyces (saccharomyces), Candida (Candida), in particular selected from the group consisting of: bifidobacterium longum (Bifidobacterium longum), Bifidobacterium lactis (Bifidobacterium lactis), Bifidobacterium animalis (Bifidobacterium animalis), Bifidobacterium breve (Bifidobacterium breve), Bifidobacterium infantis (Bifidobacterium infantis), Bifidobacterium adolescentis (Bifidobacterium adolescentis), Lactobacillus acidophilus (Lactobacillus acidophilus), Lactobacillus casei (Lactobacillus casei), Lactobacillus paracasei (Lactobacillus paracasei), Lactobacillus salivarius (Lactobacillus salivarius), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus plantarum (Lactobacillus lactis), Lactobacillus rhamnosus (Lactobacillus rhamnoides), Lactobacillus johnsonii (Lactobacillus johnsonii), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus plantarum (Lactobacillus), Lactobacillus lactis (Lactobacillus), Lactobacillus plantarum (Saccharomyces cerevisiae), Lactobacillus plantarum (Lactobacillus), Lactobacillus plantarum), Lactobacillus species (Lactobacillus), Bacillus lactis (Lactobacillus), Bacillus lactis (Bacillus lactis), Bacillus lactis (Bacillus lactis), Bacillus lactis, or mixtures of the following preferably selected from the group: bifidobacterium longum NCC3001(ATCC BAA-999), Bifidobacterium longum NCC2705(CNCMI-2618), Bifidobacterium longum NCC490(CNCMI-2170), Bifidobacterium lactis NCC2818(CNCM I-3446), Bifidobacterium breve strain A, Lactobacillus paracasei NCC2461(CNCM I-2116), Lactobacillus johnsonii NCC533(CNCM I-1225), Lactobacillus rhamnosus GG (ATCC53103), Lactobacillus rhamnosus NCC4007(CGMCC 1.3724), enterococcus faecium SF 68(NCC2768, NCIMB10415), and combinations thereof.

12. Use of a nutritional composition according to any of the preceding claims, wherein the mammal is a human, a pet animal, a cat or a dog, preferably wherein the mammal is a young mammal, more preferably an infant or a young child.

13. Use of the nutritional composition according to any of the preceding claims, wherein the composition is an infant formula, a follow-on formula, a human milk fortifier, a growing-up milk, or a pet food or supplement.

14. Human Milk Oligosaccharides (HMOs) for improving, enhancing, promoting or modulating gabaergic function in the CNS in an individual in need thereof, preferably a human infant or young child born between the ages of 7.

15. Nutritional composition comprising Human Milk Oligosaccharides (HMOs) for improving, enhancing, promoting or modulating gabaergic function in the CNS in an individual in need thereof, preferably a human infant or young child between birth and 7 years old.

16. Use of a Human Milk Oligosaccharide (HMO) for the manufacture of a nutritional composition for improving, enhancing, promoting or modulating gabaergic function in the CNS in an individual in need thereof, preferably a human infant or young child born between the ages of 7.